A sensitive assay for the quantification of glucose and lactate in the human cornea using a modified bioluminescence technique.
BACKGROUND
Quantification of glucose and lactate concentrations in human corneal extracts has been performed using spectrophotometry. We employed a bioluminescence technique to obtain a more sensitive assay for glucose and lactate and to reduce the volume of the test sample.


METHODS
The NAD(P)H bioluminescence assay (Boehringer Mannheim, Germany) was modified for glucose and lactate. Standard curves were established using a standard solution with 0.004 mM and 0.01 mM concentrations of glucose and lactate, respectively.


RESULTS
Linear standard curves ranging from 0 to 200 pmol for glucose and from 0 to 250 pmol for lactate were established. The sample volume was reduced from 100 microliters to 25 microliters compared with spectrophotometry.


CONCLUSIONS
The modified bioluminescence technique provides a highly sensitive quantification of glucose and lactate in the human cornea and thus reveals more details of the overall metabolic status of the tissue.
Frantz
A
A
Department of Ophthalmology, RWTH Aachen, Germany.
Salla
S
S
Redbrake
C
C
D000368:Aged; D002102:Cadaver; D003315:Cornea / Q000737:chemistry*; D005947:Glucose / Q000032:analysis*; D006801:Humans; D019344:Lactic Acid / Q000032:analysis*; D008163:Luminescent Measurements; D012680:Sensitivity and Specificity; D013053:Spectrophotometry; D014019:Tissue Donors
D019344:Lactic Acid; D005947:Glucose